The biological significance of the Ras oncogene, and the role of both Ras and the enzyme known as farnesyl protein transferase in the conversion of normal cells to cancer cells, are described in PCT International Publication Nos. WO95/00497 and WO95/10516. Each of those publications also describes a distinct class of compounds which inhibit the activity of the enzyme farnesyl protein transferase, and thereby the farnesylation of the Ras protein.
PCT International Publication No. WO95/10516 relates to tricyclic amide and urea compounds of the general formula (1.0) 
and their use in a method for inhibiting Ras function and the abnormal growth of cells. A number of sub-generic classes of compounds of formula (1.0) are described, which include compounds of the formulae (5.0c), (5.1c) and (5.2a) 
as well as the 11-R-isomer and 11-S-isomers of compounds (5.0c) and (5.1c). A number of specific compounds within each such sub-genus are also described therein, as is the biological activity of those compounds.
The present invention provides novel tricyclic amide compounds selected from the group consisting of: 
or pharmaceutically acceptable salts or solvates thereof.
Optical rotation of the compounds ((+)xe2x88x92or  n(xe2x88x92)xe2x88x92) are measured in methanol or ethanol at 25xc2x0 C.
This invention includes the above compounds in the amorphous state or in the cyrstalline state.
Thus, compounds of this invention include compounds selected from the group consisting of: Compounds 1.0, 2.0, 3.0, 4.0, 5.0, 7.0 and 6.0, or pharmaceutically acceptable salts thereof, wherein said compounds are as defined above.
Compounds of this invention also include compounds selected from the group consisting of: Compounds 10.0, 11.0, 12.0, 13.0, 16.0, 17.0, 18.0, 19.0, 20.0, 21.0, and 22.0, or pharmaceutically acceptable salts thereof, and wherein said compounds are as defined above.
Compounds of this invention also include compounds selected from the group consisting of: Compounds 8.0, 9.0, 14.0, and 15.0, or pharmaceutically acceptable salts thereof, and wherein said compounds are as defined above.
Compounds of this invention also include compounds selected from the group consisting of: Compounds 23.0, 25.0, 26.0, 27.0, 28.0, 29.0, 30.0, 31.0, 32.0, 33.0, 34.0, 60.0, 61.0, 62.0, 63.0, and 64.0, or pharmaceutically acceptable salts thereof, and wherein said compounds are as defined above.
Compounds of this invention also include compounds selected from the group consisting of: Compounds 23.0A, 24.0, 35.0, 36.0, 37.0, 38.0, 39.0, 40.0, 41.0, 42.0, 47.0, 48.0, 49.0, 50.0, 51.0, 52.0, 53.0, 54.0, 55.0, 56.0, 57.0, 58.0, 59.0, and 65.0, or pharmaceutically acceptable salts thereof, and wherein said compounds are as defined above.
Compounds of this invention also include compounds selected from the group consisting of: Compounds 43.0, 44.0, 45.0 and 46.0, or pharmaceutically acceptable salts thereof, and wherein said compounds are as defined above.
The preferred compounds include Compounds 5.0, 7.0, 25.0, 27.0, 29.0, and 34.0.
The preferred compounds also include Compounds 51.0 and 53.0
The preferred compounds also include Compounds 40.0 and 42.0.
More preferred compounds are Compounds 25.0, 27.0, 51.0 and 53.0.
Those skilled in the art will appreciate that the tricyclic ring system is numbered: 
Those skilled in the art will also appreciate that the S and R stereochemistry at the C-11 bond are: 
Inhibition of farnesyl protein transferase by the tricyclic compounds of this invention has not been reported previously. Thus, this invention provides a method for inhibiting farnesyl protein transferase using tricyclic compounds of this invention which: (i) potently inhibit farnesyl protein transferase, but not geranylgeranyl protein transferase I, in vitro; (ii) block the phenotypic change induced by a form of transforming Ras which is a farnesyl acceptor but not by a form of transforming Ras engineered to be a geranylgeranyl acceptor; (iii) block intracellular processing of Ras which is a farnesyl acceptor but not of Ras engineered to be a geranylgeranyl acceptor; and (iv) block abnormal cell growth in culture induced by transforming Ras.
This invention provides a method for inhibiting or treating the abnormal growth of cells, including transformed cells, by administering an effective amount of a compound of this invention. Abnormal growth of cells refers to cell growth independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the abnormal growth of: (1) tumor cells (tumors) expressing an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated as a result of oncogenic mutation in another gene; and (3) benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs.
This invention also provides a method for inhibiting or treating tumor growth (cancer) by administering an effective amount of the tricyclic compounds, described herein, to a mammal (e.g., a human) in need of such treatment. In particular, this invention provides a method for inhibiting or treating the growth of tumors expressing an activated Ras oncogene by the administration of an effective amount of the above described compounds. Examples of tumors which may be inhibited or treated include, but are not limited to, lung cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g., pancreatic carcinoma such as, for example, exocrine pancreatic carcinoma), colon cancers (e.g., colorectal carcinomas, such as, for example, colon adenocarcinoma and colon adenoma), myeloid leukemias (for example, acute myelogenous leukemia (AML)), thyroid follicular cancer, myelodysplastic syndrome (MDS), bladder carcinoma, epidermal carcinoma, breast cancers and prostate cancers.
It is believed that this invention also provides a method for inhibiting proliferative diseases, both benign and malignant, wherein Ras proteins are aberrantly activated as a result of oncogenic mutation in other genesxe2x80x94i.e., the Ras gene itself is not activated by mutation to an oncogenic formxe2x80x94with said inhibition being accomplished by the administration of an effective amount of the tricyclic compounds described herein, to a mammal (e.g., a human) in need of such treatment. For example, the benign proliferative disorder neurofibromatosis, or tumors in which Ras is activated due to mutation or overexpression of tyrosine kinase oncogenes (e.g., neu, src, abl, lck, and fyn), may be inhibited by the tricyclic compounds described herein.
The compounds of this invention inhibit farnesyl protein transferase and the farnesylation of the oncogene protein Ras. This invention further provides a method of inhibiting ras farnesyl protein transferase, in mammals, especially humans, by the administration of an effective amount of the tricyclic compounds described above. The administration of the compounds of this invention to patients, to inhibit farnesyl protein transferase, is useful in the treatment of the cancers described above.
The tricyclic compounds useful in the methods of this invention inhibit the abnormal growth of cells. Without wishing to be bound by theory, it is believed that these compounds may function through the inhibition of G-protein function, such as ras p21, by blocking G-protein isoprenylation, thus making them useful in the treatment of proliferative diseases such as tumor growth and cancer. Without wishing to be bound by theory, it is believed that these compounds inhibit ras farnesyl protein transferase, and thus show antiproliferative activity against ras transformed cells.
As used herein, the following terms are used as defined below unless otherwise indicated:
M+-represents the molecular ion of the molecule in the mass spectrum;
MH+-represents the molecular ion plus hydrogen of the molecule in the mass spectrum;
Pyridyl N-oxides are herein represented by the group 
The following solvents and reagents are referred to herein by the abbreviations indicated: tetrahydrofuran (THF); ethanol (EtOH); methanol (MeOH); acetic acid (HOAc or AcOH); ethyl acetate (EtOAc); N,N-dimethylformamide (DMF); trifluoroacetic acid (TFA); trifluoroacetic anhydride (TFAA); 1-hydroxy-benzotriazole (HOBT); m-chloroperbenzoic acid (MCPBA); triethylamine (Et3N); diethyl ether (Et2O); ethyl chloroformate ClCO2Et); 1-(3-dimethylaminopropyl)-3-ethyl carbodiimde hydrochloride (DEC); diisobutylaluminum hydride (DIBAL); isopropanol (iPrOH); dimethylsulfoxide (DMSO)
Certain compounds of the present invention may exist in different isomeric forms (e.g., enantiomers or diastereoisomers) including atropisomers (i.e., compounds wherein the 7-membered ring is in a fixed conformation such that the 11-carbon atom is positioned above or below the plane of the fused beznene rings due to the presence of a 10-bromo substituent). The invention contemplates all such isomers both in pure form and in admixture, including racemic mixtures. Enol forms are also included.
Certain basic tricyclic compounds also form pharmaceutically acceptable salts, e.g., acid addition salts. For example, the pyrido-nitrogen atoms may form salts with strong acids. Examples of suitable acids for salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methanesulfonic and other mineral and carboxylic acids well known to those in the art. The salts are prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt in the conventional manner. The free base forms may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free base forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acid and base salts are otherwise equivalent to their respective free base forms for purposes of the invention.
All such salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.
The compounds of the present invention can be prepared by the procedures described below.
The compounds of the invention having a piperidine ring (Ring IV): 
can be prepared, by techniques well known in the art, from the corresponding unoxidized pyridyl compounds: 
Thus, the compounds of the invention can be prepared from: 
The piperidine compounds (Formula I) of the invention can be prepared from the above pyridyl compounds by oxidation with meta-chloroperoxybenzoic acid. This reaction is conducted in a suitable organic solvent, e.g., dichloromethane (usually anhydrous) or methylene chloride, at a suitable temperature, to produce the compounds of the invention having the Nxe2x80x94O substituent at position 1 of Ring I of the tricyclic ring system.
Generally, the organic solvent solution of the starting tricyclic reactant is cooled to about 0xc2x0 C. before the m-chloro-peroxybenzoic acid is added. The reaction is then allowed to warm to room temperature during the reaction period. The desired product can be recovered by standard separation means. For example, the reaction mixture can be washed with an aqueous solution of a suitable base, e.g., saturated sodium bicarbonate or NaOH (e.g., 1N NaOH), and then dried over anhydrous magnesium sulfate. The solution containing the product can be concentrated in vacuo. The product can be purified by standard means, e.g., by chromatography using silica gel (e.g., flash column chromatography).
Alternatively, the piperidine compounds (Formula I) of the invention can be made from intermediate compounds of Formulas 1.1 to 65.1 using the oxidation procedure with m-chloroperoxybenzoic acid. The oxidized intermediate compounds are then reacted to produce the compounds of the invention by methods known in the art. For example, the 3,8-dihalo compounds can be produced from the intermediate: 
which is made by oxidizing the pyridyl compound 
with m-chloroperoxybenzoic acid.
The 3,7,8-trihalo compounds, 3,8,10-trihalo compounds, 3,8-dihalo compounds and the 3,10-dihalo compounds can be produced from the intermediates 
respectively.
Compounds III to VI can be prepared using the above oxidation procedure with m-chloro-peroxybenzoic acid and the pyridyl compounds 
respectively, to produce the compounds 
respectively. Compounds XI to XIV can then be converted to Compounds III to VI, respectively, by methods well known in the art.
In the above compounds the dotted line ( - - - ) represents an optional bond, and X represents CH when the optional bond is absent, and when the optional bond is present X represents C. The Nxe2x80x94O intermediates are then reacted further to produce the compounds of the invention.
Those skilled in the art will appreciate that the oxidation reaction can be conducted on racemic mixtures and the isomers can then be separated by know techniques, or the isomers can be separated first and then oxidized to the corresponding N-oxide.
Those skilled in the art will appreciate that it is preferable to avoid an excess of m-chloroperoxybenzoic acid when the oxidation reaction is carried out on the compounds having a C-11 double bond to piperidine Ring IV (e.g., compounds 5.1, 6.1, 9.1, and the like). In these reactions an excess of m-chloro-peroxybenzoic acid can cause epoxidation of the C-11 double bond.
Intermediate compounds VII, VIII, IX and X are prepared by methods known in the art, for example by methods disclosed in WO 95/10516, in U.S. Pat. No. 5,151,423 and those described below. For example, Compounds VII to X can be prepared by reacting compounds 
respectively, with C2H5OCOCl and Et3N in an inert solvent (e.g., CH2Cl2).
Intermediate Compounds XV, XVI, XVII and XVIII wherein the C-3 postion of the pyridine ring in the tricyclic structure is substituted by bromo can also be prepared by a procedure comprising the following steps:
(a) reacting an amide of the formula 
xe2x80x83wherein R11a is Br, R5a is hydrogen and R6a is C1-C6 alkyl, aryl or heteroaryl; R5a is C1-C6 alkyl, aryl or heteroaryl and R6a is hydrogen; R5a and R6a are independently selected from the group consisting of C1-6 alkyl and aryl; or R5a and R6a, together with the nitrogen to which they are attached, form a ring comprising 4 to 6 carbon atoms or comprising 3 to 5 carbon atoms and one hetero moiety selected from the group consisting of xe2x80x94Oxe2x80x94 and xe2x80x94NR9axe2x80x94, wherein R9a is H, C1-C6 alkyl or phenyl;
with a compound of the formula 
xe2x80x83wherein R1a, R2a, R3a and R4a are are independently selected from the group consisting of hydrogen and halo and R7a is Cl or Br, in the presence of a strong base to obtain a compound of the formula 
(b) reacting a compound of step (a) with
(i) POCl3 to obtain a cyano compound of the formula 
(ii) DIBALH to obtain an aldehyde of the formula 
(c) reacting the cyano compound or the aldehyde with a piperidine derivative of the formula 
xe2x80x83wherein L is a leaving group selected from the group consisting of Cl and Br, to obtain a ketone or an alcohol of the formula below, respectively: 
(d)(i) cyclizing the ketone with CF3SO3H to obtain a compound of the formula: 
xe2x80x83wherein the dotted line represents a double bond; or
(d)(ii) cyclizing the alcohol with polyphosphoric acid to obtain an Intermediate compound wherein the dotted line represents a single bond.
Methods for preparing the Intermediate compounds disclosed in WO 95/10516, U.S. Pat. No. 5,151,423 and described below employ a tricyclic ketone intermediate. Such intermediates of the formula 
wherein R11b, R1a, R2a, R3a and R4a are independently selected from the group consisting of hydrogen and halo, can be prepared by the following process comprising:
(a) reacting a compound of the formula 
(i) with an amine of the formula NHR5aR6a, wherein R5a and R6a are as defined in the process above; in the presence of a palladium catalyst and carbon monoxide to obtain an amide of the formula: 
(ii) with an alcohol of the formula R10aOH, wherein R10a is C1-C6 lower alkyl or C3-C6 cycloalkyl, in the presence of a palladium catalyst and carbon monoxide to obtain the ester of the formula 
xe2x80x83followed by reacting the ester with an amine of formula NHR5aR6a to obtain the amide;
(b) reacting the amide with an iodo-substituted benzyl compound of the formula 
xe2x80x83wherein R1a, R2a, R3a, R4a and R7a are as defined above, in the presence of a strong base to obtain a compound of the formula 
(c) cyclizing a compound of step (b) with a reagent of the formula R8aMgL, wherein R8a is C1-C8 alkyl, aryl or heteroaryl and L is Br or Cl, provided that prior to cyclization, compounds wherein R5a or R6a is hydrogen are reacted with a suitable N-protecting group.
(+)-Isomers of compounds of Formula XVI 
wherein X is CH can be prepared with high enantioselectivity by using a process comprising enzyme catalyzed transesterification. Preferably, a racemic compound of Formula XVI, wherein X is C and the double bond is present, is reacted with an enzyme such as Toyobo LIP-300 and an acylating agent such as trifluoroethly isobutyrate; the resultant (+)-amide is then hydrolyzed, for example by refluxing with an acid such as H2SO4, to obtain the corresponding optically enriched (+)-isomer wherein X is CH. Alternatively, a racemic compound of Formula XVI, wherein X is C and the double bond is present, is first reduced to the corresponding racemic compound of Formula XVI wherein X is CH and then treated with the enzyme (Toyobo LIP-300) and acylating agent as described above to obtain the (+)-amide, which is hydrolyzed to obtain the optically enriched (+)-isomer.
The compound of Preparative Example 21 is obtained in the crystalline state. Those skilled in the art will appreciate that compounds obtained in the amorphous state can be obtained in the crystalline state by crystallizing the amorphous materials from solvents or solvent mixtures such as acetone, diethyl ether, ethyl acetate, ethanol, 2-propanol, tert-butyl ether, water and the like according to procedures well known in the art.
Those skilled in the art will also appreciate that the racemic mixture of Compound 11.0 can be made according to the procedures described below. For Example, the intermediate of Preparative Example 6 can be used to prepare Compound 11.0.
Compounds of the invention having a piperazine ring 
can be prepared from the tricyclic ketone: 
Ketone XX can be prepared by oxidation of the corresponding pyridyl compound: 
with m-chloroperoxybenzoic acid.
Ketone XX can be converted to the corresponding C-11 hydroxy compound which in turn can be converted to the corresponding C-11 chloro compound 
Compound XXIII can then be reacted with piperazine to produce the intermediate: 
Intermediate XXIV can then be reacted with the reagents which will provide the desired final product.
The above reactions are well known in the art and are illustrated in the examples below.